The objective of this study was to identify allelic variations at $Glu-1$ loci of wheat ($Triticum$ $aestivum$ L.) advanced lines derived from hybridization of bread wheat and synthetic hexaploid wheats (2n = 6x = 42; AABBDD). Locally adapted wheat genotypes were crossed with synthetic hexaploid wheats. From the 134 different cross combinations made, 202 $F_8$ advanced lines were selected and their HMW-GS composition was studied using SDS-PAGE. In total, 24 allelic variants and 68 HMW-GS combinations were observed at $Glu-A1$, $Glu-B1$, and $Glu-D1$ loci. In bread wheat, the $Glu-D1$ locus is usually characterized by subunits 1Dx2+1Dy12 and 1Dx5+1Dy10 with the latter having a stronger effect on bread-making quality. The subunit 1Dx5+1Dy10 was predominantly observed in these advanced lines. The inferior subunit 1Dx2+1Dy12, predominant in adapted wheat germplasm showed a comparative low frequency in the derived advanced breeding lines. Its successful replacement is due to the other better allelic variants at the $Glu-D1$ locus inherited in these synthetic hexaploid wheats from $Aegilops$ $tauschii$ (2n = 2x = 14; DD).